Chemotherapy resistance research of lung cancer based on micro-fluidic chip system with flow medium

Abstract

Micro total analysis systems (-TAS) or labs-on-achip, have been spreading rapidly due to their desirable characteristics, including reductions in reagent consumption, space requirements and analysis time. This work aimed at establishing an integrated microfluidic system which can supply the cells with fresh medium of oxygen and nutrition continuously at a control flow rate mimicking the microenvironment in vivo. Human non-small cell lung cancer cell line SPCA1 was seeded in a microchip supplied with fresh medium at a constant rate of 15 mm/24 h controlled by a pump. The expression of P-gp for verapamil-pretreated or non-pretreated cells was assayed with immunofluorescence. Both groups cells were exposed to anticancer drug VP-16 at 30 microM for 6 h before the apoptosis analysis online. The results indicated that the cells could grow and spread well for 4 days in the microfluidic system successively furnished with fresh medium. Immunofluorescence assay showed that the intensity of the fluorescence for the verapamil-pretreated cells was obvious weak compared with that of nonpretreated cells. Apoptosis analysis demonstrated that the percentage of apoptotic cells for verapamil-pretreated group increased around twofold compared with that of nonverapamil pretreated group (26.5+/-2.5% versus 10.9+/- 0.85%, p<0.05), showing a similar results as by flow cytometry analysis. All these indicate that P-gp plays an important role in the resistance to VP-16 in SPCA1, the microfluidic system provides a suitable environment for cells survival and is valuable in long time cell culture and bioassays mimicking the microenvironment in vivo and deserved to be studied further.

Biomimetic tissues on a chip for drug discovery

“…A second engineering challenge is to expose different combinations or concentrations of drugs to cells cultured on-chip. Two groups recently accomplished this step by exposing human lung cancer cells to different concentrations of the anticancer drug verapamil (VP-16) [84,85]. In one study, the percentage of apoptotic cells in the verapamil-pretreated group was approximately double that of the control group, in agreement with previous flow cytometry analysis [85].…”

Development of a double‐layer microfluidic chip with flow medium for chemotherapy resistance analysis of lung cancer

“…But shear stress caused by fluid velocity is bad for cell growth, so fluid velocity during perfusion of cell culture is relatively low and is dictated by the balance between adequate mass transport and limited hydrodynamic effects [27]. Compared with our previous single layer chip [19], the new double-layer one has some advantages such as the possibility of the injection of the cells with medium and the drugs through different ports (inlet A for cells, inlets B and C for drugs), which does not only mean the convenience but also the reduction of the chance of contamination and mixture by the disparate mediator; and the possibility to make sure that two groups of cells were in the same condition before further culture and analysis in parallel. With this device, human lung cancer cell line SPCA-1 grew and propagated perfectly for at least 96 h. The designed double-layer chip made sure that the multioperations including preparation and separation of SPCA-1cells, identification of MRP1 protein, addition of the drug of VP-16 or dyes and detection of cell viability integrated onto the platform systematically and conveniently, fully exploiting the integrated advantage of microfluidic technology over the conventional platform in terms of time and reagent consumption as well as sensitivity.…”

“…The PDMS microchip was fabricated by replica molding PDMS against the masters, and the resulting PDMS microchip was irreversibly bonded to a glass slide assisted by oxygen plasma surface treatment (150 mTorr, 50 W, 20 s). The upper layer of the double-layer chip was mainly used to supply the cells with flow fresh medium of oxygen and nutrition through an MS26 injection pump mimicking the microenvironment in vivo, with accurately controlled rate of 10 mm/24 h as we previously reported [19]. It can deliver 2 mL fluid by one pulse of the motor.…”

Organ-on-a-Chip Platforms for Drug Delivery and Cell Characterization: A Review

“…Investigating cell-cell interactions, cell-blood flow, and cell-gas flow in the respiratory tract is essential for both physiological research and drug delivery (58) tests. Some microfluidic platforms are designed to culture lung cancer cells and screen their response to drugs (61) and electrical stimulation. (62) However, these platforms do not mimic both the deformation of the lung tissue during breathing and the bi-cell barriers between blood and air.…”